UNIVERSITY OF EXETER
Risk Assessment Of Genetically Modified Micro-organisms Where The Primary Risk Posed Is To The Environment Rather Than To Human Health
This risk assessment form is for use in cases where the primary risk posed by the Genetically Modified (GM) micro-organism is to the environment rather than to human health. Please obtain an alternative form if the primary risk posed by the GM micro-organism is to human health.
Part 1 of this form should be completed for all projects. Parts 2 and 3 are for risk assessments where the issues involved are complex and require detailed consideration.
Please type in the boxes - you can replace the notes in the boxes.
Provide justification, explanation and detail in all parts of this form.
Carry out your risk assessment in accordance with Section 2 of the ACGM Compendium of Guidance, particularly Section 2A. These are accessible on the Biology fileserver in: G:\Safety\Genetic Modification\ACGM Compendium of Guidance\
This risk assessments should be reviewed whenever there is a significant change in the activity involving GM.
Part 1
(a) Title of project and principal investigator
(b) Scientific goals of project
This information provides a useful background and puts the work in context. This step is not obligatory. The scientific goals need not be disclosed if presenting the information presents problems in relation to intellectual property rights or commercial sensitivity.
(c) An overview of the different types of GMM that will be constructed
This overview should consist of 1 or 2 paragraphs, outlining the scope of the project and setting the boundaries of exactly what work will be done. This overview should be complemented with lists of recipient strains, vectors and inserts that will be used in the project.
(i) List of recipient strain(s)
Cover the name of the strain, the name of the wild-type organism from which it is derived and the extent to which it is disabled.
(ii) List of vector(s)
Cover names and any diasabling mutations.
(iii) List of function of inserted gene(s)
In doing this genes should be identified in such a way that an outside reviewer will have a general idea of their function i.e. providing a three-letter name may not be sufficient. Where the function of a gene is unknown, it may help to provide details of any known homologues.
(d) An indication of the most hazardous GMM
Considering both human health and the environment the most hazardous GMM that will be constructed in this work should be identified. This will be the most hazardous combination of recipient strain, vector and inserted material from the lists made under part (c). With some projects it will not be clear that any one GMM will be any more hazardous than any of the others (e.g. if all the work is class 1). If this is the case this should be stated.
(e) Are you confident that for all of the GMMs covered by this assessment there are no harmful properties associated with the recipient strain, the vector, or the inserted material?
If the answer to this question is ‘No’ or you are in any way unsure, Part 2 of this form should be completed.
(f) Are you confident that none of the final GMMs could be hazardous to humans or the environment?
If the answer to this question is ‘No’ or you are in any way unsure, Part 2 of this form should be completed.
If the GMM meets BOTH of the above criteria, you may believe that you have sufficient information at this stage to classify the project to class 1, as defined in the Contained Use Regulations 2000. In order to do this you should be confident that even in the event of a total breach of containment the genetically modified organism would be of no or negligible risk to human health or the environment.
If you are assigning the work to class 1 according to the above streamlined procedures, sign the form below and submit it to the GM Safety Officer (GMSO) who will arrange for it to be considered by the local Genetic Modification Safety Committee (GMSC). Work should not be commenced until this form has been reviewed by the GMSC.
If you have any uncertainty as to whether the proposal meets the above criteria you should undertake a more detailed risk assessment by completing Part 2 and Part 3 of this form.
Signature of Proposer:Date:
Background notes
It is not appropriate to consider non-disabled pathogens of plants, humans, animals or insects as inherently safe recipient micro-organisms. Examples of inherently safe recipient micro-organisms which, depending on the nature of the insert, would in most cases be expected to form the basis of extremely safe GMMs are as follows:
(i) Escherichia coli K12, Saccharomyces cerevisiae, Schizosaccharomyces pombe
(ii) Defective retrovirus produced from packaging line in which the helper genes are located in two separate blocks of DNA (thus eliminating the possibility of a reversion to replication competence by a single recombination event) and which are self-inactivated to prevent insertional activation.
(iii) E1 deleted adenovirus
The types of gene which when cloned into particular recipient micro-organisms might give rise to a harmful phenotype can be divided into two types. First the gene might encode a product that could act directly to cause harmful effects e.g. a toxin gene or a cytokine. Second the gene might encode a product that could act alongside the exis ting characteristics of the recipient micro-organism so as to endow the GMM with altered pathogenic properties e.g. a pathogenicity gene or an engineered viral envelope gene with an altered receptor binding capacity.
Part 2 – Continuation Form for Use in Cases Where a Detailed Risk Assessment is Necessary
Projects that have a broad scope will involve the construction of several GMMs. Part 2 is designed for the detailed assessment of a single GMM. In general, as long as the scope of the work has been satisfactorily outlined in Part 1, it is sufficient to complete Part 2 for the most hazardous GMM being constructed. This will be the one identified in Part 1(d). In cases where there are two or more hazardous GMMs, with quite different properties, two or more copies of Part 2 may need to be completed.
In the format shown here, environmental issues are considered first and used as the basis for assigning provisional containment prior to addressing hazards to human health. In cases where hazards to human health are of prime concern and there is little possibility of harm to the environment e.g. work with human pathogens, please use the alternative form in which human health hazards are considered first.
(a) Identification of any hazards to the environment
(i) Hazards associated with the recipient micro-organism (e.g. bacterial host or viral vector)
Factors to consider include whether the recipient micro-organism is capable of infecting any plants, animals or insects in the environment and whether there is any possibility of any disabling mutations being complemented or reverting. In particular it should be ascertained whether the recipient micro-organism is a pathogen that is controlled by DEFRA by consulting the DEFRA list of pathogens and the scientific literature. Non-disabled animal pathogens by definition cannot be said to be of no or negligible risk hence will always be Class 2 or higher. Consider whether the micro-organism will survive in the environment (e.g. adenovirus can survive for several months extracellularly). Consider the host range of the recipient micro-organisms including domestic/cultivated, wild and endangered species.
(ii) Hazards arising directly from the inserted gene product
Factors to consider include whether the sequence encodes an insect or animal toxin, a modulator of growth or differentiation (e.g. hormone or cytokine), or a product which can cause silencing of a gene encoding a crucial metabolic enzyme in susceptible hosts.
(iii) Hazards arising from the alteration of existing traits (e.g. alteration of pathogenicity, host range or tissue tropism)
One factor to consider is whether the inserted sequence encodes a pathogenicity determinant, such as an adhesin, a penetration factor or a surface component providing resistance to host defence mechanisms. Another important consideration is whether the inserted gene encodes a surface component, envelope protein or capsid protein that might bind to a different receptor to that used by the recipient micro-organism. The impact on fitness hence competitiveness of the GMM should be considered i.e. inserted gene may endow a selective advantage e.g. resistance to desiccation, resistance to antibiotics.
(iv) The potential hazards of sequences within the GMM being transferred to related micro-organisms
Factors to consider include whether widespread dissemination of the inserted gene as a result, for example, of either gene transfer or recombination of the GMM with a wild-type micro-organism, would be a matter of concern e.g. gene associated with antibiotic resistance. If this is the case an important consideration will be whether, in the event of a breach of containment, the GMM could survive, establish and disseminate in the environment for long enough for such a gene transfer to take place. You should consider whether the vectors being used e.g. conjugative plasmids, transposable elements will contribute to gene spread. The prevalence of related micro-organisms with which gene transfer may be possible should be considered.
(b) Assignment of a provisional containment level that is adequate to protect against hazards to the environment.
This step will involve considering the containment level necessary to control the risk of the recipient micro-organism and making a judgement about whether the modification will result in a GMM which is more hazardous, less hazardous, or about the same. Sometimes it may help to compare the GMM with the relative hazard presented by other organisms. Important considerations include what is the mode of transmission of the pathogen e.g. for an animal pathogen: faecal-oral route, respiratory route or bloodborne route? You should consult the Containment Tables (e.g. Table 1a for laboratories and Table 1c for animal units) in Schedule 8 of the GM Organisms (Contained Use) Regulations 2000 to determine the appropriate containment level.
(c) Hazards to human health
(i) Hazards associated with the recipient micro-organism (e.g. bacterial host or viral vector)
Factors to consider include whether the recipient micro-organism is listed in ACDP hazard groups 2, 3 or 4. Other relevant factors may be the micro-organism’s mode of transmission, disease symptoms, host range, and tissue tropism as well as an indication as to whether vaccines or chemotherapeutic agents are available. Information should also be provided on any disabling mutations and whether there is any possibility of any disabling mutations being complemented or reverting.
(ii) Hazards arising directly from the inserted gene product (e.g. cloning of a toxin gene or oncogene)
Consideration should be given to whether the inserted DNA encodes a toxin, an oncogenic protein, an allergen, a modulator of growth or differentiation (hormone or cytokine) or any other protein which may result in potentially harmful biological activity. Where the function of the inserted gene is unknown, it may help to describe the function of any known homologues. Please note that even a normal human gene may be harmful if overexpressed, especially if the overexpression is in tissues that do not normally express the protein.
(iii) Hazards arising from the alteration of existing traits (e.g. alteration of pathogenicity, host range, tissue tropism, mode of transmission or host immune response)
One factor to consider is whether the inserted gene encodes a pathogenicity determinant, such as an adhesin, a penetration factor or a surface component providing resistance to host defence mechanisms. Another important consideration is whether the inserted gene encodes a surface component, envelope protein or capsid protein that might bind to a different receptor to that used by the recipient micro-organism. Consideration should also be given to whether the inserted DNA (or the plasmid sequence) encodes resistance to a drug or antibiotic that might be used for the treatment of a laboratory-acquired infection.
(iv) The potential hazards of sequences within the GMM being transferred to related micro-organisms
Factors to consider include whether widespread dissemination of the inserted gene as a result, for example, of either gene transfer or recombination of the GMM with a wild-type micro-organism, would be a matter of concern. If this is the case an important consideration will be whether, in the event of a breach of containment the GMM could survive in the environment for long enough for such a gene transfer to take place.
(d) Consideration of the nature of the work to be undertaken and a detailed review of the control measures
(i) Are any of the work procedures likely to generate aerosols?
If so, should the work be undertaken in a safety cabinet or isolator?
(ii) How will waste materials be disposed of?
Include both solid and liquid laboratory waste and waste from experiments with infected animals.
(iii) Will it be necessary to use sharps?
E.g. does work involve glass Pasteur pipettes?
(iv) If the work involves the experimental infection of animals is it known whether the animal will shed the GMM?
(v) If the work involves the experimental infection of plants what is known about the likely route of transmission of the GMM?
For example, is the micro-organism insect-borne or carried in run-off water? This will have important implications for the type of glasshouse used.
(vi) In the case of organisms whose multiplication involves a complex life-cycle will the work involve the propagation of organisms that are in stages in that life-cycle that are particularly hazardous?
Examples include the propagation of the infective stages of parasites or the release of spores from fungi. Consideration should be given to all potential routes of transmission including those that might not be used naturally.
(vii) Have any disinfectants been validated under the actual conditions of use?
For example, if disinfectant is being used for the treatment of virus in tissue culture medium, is it known that the disinfectant is effective in the presence of high levels of protein?
(viii) Does the nature of this work preclude it being undertaken by any workers who have a serious skin condition (e.g. eczema) or other health problems that might make them more susceptible to infection (e.g. some kind of immunological defect)?
(ix) Will workers receive any vaccinations or health surveillance? Will it be necessary to preclude workers handling animal pathogens from contact with any susceptible animal species?
(e) Consideration of whether there is a need to assign additional measures over and above the provisional level of containment.
It should be noted that the containment measures set out in Part II of Schedule 8 of the Contained Use Regulations will include some measures that are required where and to the extent that the risk assessment shows they are required
Additional measures may be necessary in any of the following circumstances:
(i) to take full account of any properties of the GMM that may be hazardous to human health.
(ii) to protect the environment.
(iii) to provide additional safeguards for particular work procedures.
Part 3. Final assignment of containment measures and risk class
The following aspects of this project are assigned to class 1.
The following aspects of this project are assigned to class 2.
The following aspects of this project are assigned to class 3.
The following aspects of this project are assigned to class 4.
Signature of Proposer:Date:
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